CA2306346A1 - Tissue-engineered tubular construct having circumferentially oriented smooth muscle cells - Google Patents

Tissue-engineered tubular construct having circumferentially oriented smooth muscle cells Download PDF

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CA2306346A1
CA2306346A1 CA002306346A CA2306346A CA2306346A1 CA 2306346 A1 CA2306346 A1 CA 2306346A1 CA 002306346 A CA002306346 A CA 002306346A CA 2306346 A CA2306346 A CA 2306346A CA 2306346 A1 CA2306346 A1 CA 2306346A1
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construct
cells
substrate
tissue
muscular
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CA2306346C (en
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Laura E. Niklason
Jinming Gao
Robert S. Langer
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Massachusetts Institute of Technology
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/507—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/06—Tubular
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/26—Constructional details, e.g. recesses, hinges flexible
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/14—Scaffolds; Matrices
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/14—Pressurized fluid
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0068—General culture methods using substrates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/069—Vascular Endothelial cells
    • C12N5/0691—Vascular smooth muscle cells; 3D culture thereof, e.g. models of blood vessels
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/20—Transition metals
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/32—Amino acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2503/00—Use of cells in diagnostics
    • C12N2503/04—Screening or testing on artificial tissues
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/30—Synthetic polymers
    • C12N2533/40—Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers

Abstract

Improved methods for the production of tissue-engineered constructs, including muscular tissue constructs such as vascular constructs, are disclosed. The methods include the use of improved substrates for cell growth, improved cell culture media for cell growth, and the use of distensible bodies to impart pulsatile stretching force to the lumens of constructs during growth. Also disclosed are improved products and methods for making those products, including substrates and cell culture media, for tissue engineering and tissue culture generally. Improved muscular tissue constructs, including vascular constructs, are also disclosed, which may be used in medicine for the repair or replacement of damaged natural structures.

Claims (80)

1. A method for producing a muscular tissue-engineered construct comprising the steps of:
(a) providing a porous substrate comprising a biocompatible material, said porous substrate having an inner surface and an outer surface; wherein said inner surface of the porous substrate defines a lumen;
(b) providing a distensible body within the lumen of said porous substrate, wherein said distensible body is capable of distending within the lumen so as to contact the inner surface of the substrate;
(c) contacting said porous substrate with a suspension comprising smooth muscle cells capable of adhering thereto, thereby forming a primary cell-seeded construct;
(d) maintaining said primary cell-seeded construct for a first growth period in an environment suitable for growth of said smooth muscle cells in the primary cell-seeded construct to form a primary tissue-engineered construct; wherein (e) during said first growth period, cyclical increases in pressure within said distensible body are provided, thereby causing said distensible body to distend within the lumen of the construct, thereby applying pulsatile stretch to the construct.
2. A method for producing a muscular tissue-engineered construct comprising the steps of (a) providing a porous substrate comprising a biocompatible material, said porous substrate having an inner surface and an outer surface; wherein said inner surface of the porous substrate defines a lumen;
(b) contacting said porous substrate with a suspension comprising smooth muscle cells capable of adhering thereto, thereby forming a primary cell-seeded construct;
(c) providing a sleeve around a portion of the exterior of said porous substrate, wherein said sleeve is capable of resisting distension of said substrate in response to pressure within the lumen of said substrate;
(d) maintaining said primary cell-seeded construct for a first growth period in an environment suitable for growth of said smooth muscle cells in the primary cell-seeded construct to form a primary tissue-engineered construct; wherein (e) during said first growth period, intralumenal flow within said lumen is provided, thereby causing said substrate to distend within the sleeve, thereby contacting said sleeve and receiving mechanical support therefrom.
3. A method as in claim 2 wherein (e) during said first growth period, cyclical increases in pressure within said lumen are provided, thereby causing said substrate to cyclically distend within the sleeve, thereby applying pulsatile stretch to the construct.
4. A method for producing a muscular tissue-engineered construct comprising the steps of:
(a) providing a porous substrate comprising a biocompatible material, said porous substrate having an inner surface and an outer surface; wherein said inner surface of the porous substrate defines a lumen, and wherein said inner surface is substantially less porous than said outer surface, and said inner surface is capable of resisting distension of said substrate in response to pressure within the lumen of said substrate;
(b) contacting said porous substrate with a suspension comprising smooth muscle cells capable of adhering thereto, thereby forming a primary cell-seeded construct;
(c) maintaining said primary cell-seeded construct for a first growth period in an environment suitable for growth of said smooth muscle cells in the primary cell-seeded construct to form a primary tissue-engineered construct; wherein (d) during said first growth period, intralumenal flow within said lumen is provided, thereby causing said substrate to distend.
5. A method as in claim 4 wherein (e) during said first growth period, cyclical increases in pressure within said lumen are provided, thereby causing said substrate to cyclically distend and applying pulsatile stretch to the construct.
6. A method as in any one of claims 1-5 wherein said porous substrate comprises a synthetic polymeric material having a hydrophilic surface.
7. A method as in any one of claims 1-5 further comprising the additional steps of:
(a) contacting said primary cell-seeded construct or said primary tissue-engineered construct with a suspension comprising a second type of mammalian cells capable of adhering thereto, thereby forming a secondary cell-seeded construct; and (b) maintaining said secondary cell-seeded construct for a second growth period in an environment suitable for growth of said second type of cells in said primary cell-seeded construct or said primary tissue-engineered construct to form a secondary tissue-engineered construct.
8. A method as in claim 7 wherein said muscular, tissue-engineered construct is a vascular tissue construct, said porous substrate is a substantially tubular substrate;
said first type of mammalian cells are smooth muscle cells;
said second type of mammalian cells are endothelial cells; and said endothelial cells are contacted with the inner surface of said lumen.
9. A method as in any one of claims 1, 3 or 5 wherein said pulsatile stretch causes an increase in an inner diameter of said construct of at least 1%.
10. A method as in claim 9 wherein said pulsatile stretch causes an increase in an inner diameter of said construct of at least 2%.
11. A method as in claim 10 wherein said pulsatile stretch causes an increase in an inner diameter of said construct of at least 6%.
12. A method for producing a tissue-engineered construct comprising the steps of:
(a) providing a substrate comprising a biocompatible synthetic polymer, said substrate having a hydrophilic surface;
(b) contacting said substrate with a suspension comprising a first type of mammalian cells capable of adhering thereto, thereby forming a primary cell-seeded construct;
and (c) maintaining said primary cell-seeded construct for a first growth period in an environment suitable for growth of said mammalian cells in the primary cell-seeded construct to form a primary tissue-engineered construct.
13. A method as in claim 12 further comprising the additional steps of:
(a) contacting said primary cell-seeded construct or said primary tissue-engineered construct with a suspension comprising a second type of mammalian cells capable of adhering thereto, thereby forming a secondary cell-seeded construct; and (b) maintaining said secondary cell-seeded construct for a second growth period in an environment suitable for growth of said second type of cells in said primary cell-seeded construct or said primary tissue-engineered construct to form a secondary tissue-engineered construct.
14. A method as in claim 6 wherein said mammalian cells are selected from the group consisting of smooth muscle cells, epithelial cells, endothelial cells, fibroblasts, myoblasts, hepatocytes, bile duct cells, pancreatic islet cells, thyroid, parathyroid, adrenal, hypothalamic, pituitary, ovarian, testicular, or salivary cells, cardiac muscle cells, renal cells, chondrocytes, nerve cells, and progenitor cells.
15. A method as in claim 7 wherein said mammalian cells are selected from the group consisting of smooth muscle cells, epithelial cells, endothelial cells, fibroblasts, myoblasts, hepatocytes, bile duct cells, pancreatic islet cells, thyroid, parathyroid, adrenal, hypothalamic, pituitary, ovarian, testicular, or salivary cells, cardiac muscle cells, renal cells, chondrocytes, nerve cells, and progenitor cells.
16. A method as in claim 12 wherein said mammalian cells are selected from the group consisting of smooth muscle cells, epithelial cells, endothelial cells, fibroblasts, myoblasts, hepatocytes, bile duct cells, pancreatic islet cells, thyroid, parathyroid, adrenal, hypothalamic, pituitary, ovarian, testicular, or salivary cells, cardiac muscle cells, renal cells, chondrocytes, nerve cells, and progenitor cells.
17. A method as in claim 13 wherein said mammalian cells are selected from the group consisting of smooth muscle cells, epithelial cells, endothelial cells, fibroblasts, myoblasts, hepatocytes, bile duct cells, pancreatic islet cells, thyroid, parathyroid, adrenal, hypothalamic, pituitary, ovarian, testicular, or salivary cells, cardiac muscle cells, renal cells, chondrocytes, nerve cells, and progenitor cells.
18. A method as in claim 12 wherein said polymeric material comprises a polymer selected from the group consisting of polyesters of hydroxycarboxylic acids, polyanhydrides of dicarboxylic acids, and copolymers of hydroxy carboxylic acids and dicarboxylic acids.
19. A method as in claim 18 wherein said polymeric material is selected from the group consisting of polymers or copolymers of glycolic acid, lactic acid, and sebacic acid.
20. A method as in claim 12 wherein said substrate comprises a porous mesh of fibers, said fibers having a diameter of between approximately 5-20 µm.
21. A method as in claim 20 wherein said fibers have a diameter of between approximately 10-15 µm.
22. A method as in claim 12 wherein said substrate comprises a porous mesh of fibers, substantially parallel fibers in said mesh being separated by approximately 20-200 µm.
23. A method as in claim 22 wherein said substantially parallel fibers in said mesh are separated by approximately 50-100 µm.
24. A method as in claim 12 wherein said substrate is characterized by a void volume of greater than 90%.
25. A method as in claim 24 wherein said substrate is characterized by a void volume of greater than 95%.
26. A method as in claim 12 wherein said substrate has an average pore size of less than 200 µm.
27. A method as in claim 26 wherein said substrate has an average pore size of less than 175 µm.
28. A method as in claim 12 wherein said substrate has an average pore size of less than 150 µm.
29. A method as in claim 12 wherein said hydrophilic surface comprises a multiplicity of hydrophilic chemical groups on said surface, selected from the group consisting of carboxyl, hydroxyl, thiol, amine, sulfonyl, guanidine, and amide groups.
30. A method as in claim 29 wherein said hydrophilic groups have a density of at least 5 pmol/cm2.
31. A method as in claim 29 wherein said hydrophilic groups have a density of at least 10 pmol/cm2.
32. A method as in claim 29 wherein said hydrophilic groups have a density between 5 and 20 pmol/cm2.
33. A method as in claim 12 wherein said hydrophilic surface has a contact angle of less than 20°.
34. A method as in claim 33 wherein said hydrophilic surface has a contact angle of less than 15°.
35. A method as in claim 33 wherein said hydrophilic surface has a contact angle of less than 10°.
36. A method as in claim 33 wherein said hydrophilic surface has a contact angle of less than 5°.
37. A method as in claim 6 wherein said mammalian cells are smooth muscle cells and said environment in said first growth period comprises a standard cell culture medium supplemented with about 0.01-0.1 g/L of at least one amino acid selected from the group consisting of proline, glycine, and alanine.
38. A method as in claim 37 wherein said mammalian cells are smooth muscle cells and said environment in said first growth period comprises a standard cell culture medium supplemented with about 0.02-0.06 g/L of at least one amino acid selected from the group consisting of proline, glycine, and alanine.
39. A method as in claim 6 wherein said mammalian cells are smooth muscle cells and said environment in said first growth period comprises a standard cell culture medium supplemented with about 0.01-0.1 g/L of vitamin C.
40. A method as in claim 39 wherein said mammalian cells are smooth muscle cells and said environment in said first growth period comprises a standard cell culture medium supplemented with about 0.02-0.06 g/L of vitamin C.
41. A method as in claim 6 wherein said mammalian cells are smooth muscle cells and said environment in said first growth period comprises a standard cell culture medium supplemented with about 0.5-5.0 µg/L of a copper salt.
42. A method as in claim 40 wherein said mammalian cells are smooth muscle cells and said environment in said first growth period comprises a standard cell culture medium supplemented with about 1.0-3.0 µg/L of a copper salt.
43. A substrate for use in tissue culture comprising a three-dimensional scaffold of a biocompatible synthetic polymer having a hydrophilic surface.
44. A method as in claim 43 wherein said polymeric material comprises a polymer selected from the group consisting of polyesters of hydroxycarboxylic acids, polyanhydrides of dicarboxylic acids, and copolymers of hydroxy carboxylic acids and dicarboxylic acids.
45. A method as in claim 44 wherein said polymeric material is selected from the group consisting of polymers or copolymers of glycolic acid, lactic acid, and sebacic acid.
46. A method as in claim 43 wherein said substrate comprises a porous mesh of fibers, said fibers having a diameter of between approximately 5-20 µm.
47. A method as in claim 46 wherein said fibers have a diameter of between approximately 10-15 µm.
48. A method as in claim 43 wherein said substrate comprises a porous mesh of fibers, substantially parallel fibers in said mesh being separated by approximately 20-200 µm.
49. A method as in claim 48 wherein said substantially parallel fibers in said mesh are separated by approximately 50-100 µm.
50. A method as in claim 43 wherein said substrate is characterized by a void volume of greater than 90%.
51. A method as in claim 50 wherein said substrate is characterized by a void volume of greater than 95%.
52. A method as in claim 43 wherein said substrate has an average pore size of less than 200 µm.
53. A method as in claim 52 wherein said substrate has an average pore size of less than 175 µm.
54. A method as in claim 52 wherein said substrate has an average pore size of less than 150 µm.
55. A method as in claim 43 wherein said hydrophilic surface comprises a multiplicity of hydrophilic chemical groups on said surface, selected from the group consisting of carboxyl, hydroxyl, thiol, amine, sulfonyl, guanidine, and amide groups.
56. A method as in claim 55 wherein said hydrophilic groups have a density of at least 5 pmol/cm2.
57. A method as in claim 55 wherein said hydrophilic groups have a density of at least 10 pmol/cm2.
58. A method as in claim 55 wherein said hydrophilic groups have a density between 5 and 20 pmol/cm2.
59. A method as in claim 43 wherein said hydrophilic surface has a contact angle of less than 20°.
60. A method as in claim 59 wherein said hydrophilic surface has a contact angle of less than 15°.
61. A method as in claim 59 wherein said hydrophilic surface has a contact angle of less than 10°.
62. A method as in claim 59 wherein said hydrophilic surface has a contact angle of less than 5°.
63. A substrate as in claim 43 wherein said substrate comprises a multiplicity of polyester or polyanhydride bonds and said hydrophilic surface is formed by at least partial hydrolysis of said bonds at said surface.
64. A muscular, tubular tissue-engineered construct comprising a substantially tubular construct of living mammalian tissue having a first end and a second end, an inner surface and an outer surface; wherein the first end, the second end, and the inner surface of the construct define a lumen passing through the construct; and wherein tissue between said inner surface and said outer surface defines a wall of said construct;
wherein said wall comprises mammalian smooth muscle cells oriented circumferentially about said lumen.
65. A muscular, tubular tissue-engineered construct as in claim 64 wherein wherein said mammalian smooth muscle cells in said wall have a cell density of at least 10 7 cells/cc.
66. A muscular, tubular tissue-engineered construct as in claim 65 wherein wherein said mammalian smooth muscle cells in said wall have a cell density of at least 10 8 cells/cc.
67. A muscular, tubular tissue-engineered construct as in claim 64 wherein wherein said tubular construct is capable of withstanding an internal pressure of at least 100 mm Hg for a sustained period without rupturing.
68. A muscular, tubular tissue-engineered construct as in claim 67 wherein wherein said tubular construct is capable of withstanding an internal pressure of at least 110 mm Hg for a sustained period without rupturing.
69. A muscular, tubular tissue-engineered construct as in claim 67 wherein wherein said tubular construct is capable of withstanding an internal pressure of at least 120 mm Hg for a sustained period without rupturing.
70. A muscular, tubular tissue-engineered construct as in claim 67 wherein wherein said tubular construct is capable of withstanding an internal pressure of at least 130 mm Hg for a sustained period without rupturing.
71. A muscular, tubular tissue-engineered construct as in claim 64 wherein wherein said tubular construct is capable of withstanding an internal shear force of at least dynes/cm2 for a sustained period without rupturing.
72. A muscular, tubular tissue-engineered construct as in claim 71 wherein wherein said tubular construct is capable of withstanding an internal shear force of at least dynes/cm2 for a sustained period without rupturing.
73. A muscular, tubular tissue-engineered construct as in claim 71 wherein wherein said tubular construct is capable of withstanding an internal shear force of at least dynes/cm2 for a sustained period without rupturing.
74. A muscular, tubular tissue-engineered construct as in claim 71 wherein wherein said tubular construct is capable of withstanding an internal shear force of at least dynes/cm2 for a sustained period without rupturing.
75. A muscular, tubular tissue-engineered construct as in claim 64 wherein said wall further comprises a biocompatible synthetic polymeric material.
76. A muscular, tubular tissue-engineered construct as in claim 64 wherein said outer surface is substantially free of an adventitia.
77. A muscular, tubular tissue-engineered construct as in claim 64 wherein said wall is substantially free of an intermediate layer of an intima.
78. A muscular, tubular tissue-engineered construct as in claim 64 wherein said wall is substantially free of an internal elastic lamina of an intima.
79. A muscular, tubular tissue-engineered construct as in claim 64 wherein said wall is substantially free of fibroblasts in an intimal layer.
80. A muscular, tubular tissue-engineered construct as in claim 64 wherein said wall is substantially free of fibroblasts in a medial layer.
CA2306346A 1997-07-03 1998-07-02 Tissue-engineered tubular construct having circumferentially oriented smooth muscle cells Expired - Lifetime CA2306346C (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US5163497P 1997-07-03 1997-07-03
US60/051,634 1997-07-03
US5255397P 1997-07-15 1997-07-15
US60/052,553 1997-07-15
PCT/US1998/013828 WO1999001538A1 (en) 1997-07-03 1998-07-02 Tissue-engineered constructs

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CA2306346A1 true CA2306346A1 (en) 1999-01-14
CA2306346C CA2306346C (en) 2010-09-14

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EP (2) EP1019492A1 (en)
CA (1) CA2306346C (en)
WO (1) WO1999001538A1 (en)

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US8143055B2 (en) 2008-06-24 2012-03-27 The Curators Of The University Of Missouri Self-assembling multicellular bodies and methods of producing a three-dimensional biological structure using the same
US8747880B2 (en) 2010-02-02 2014-06-10 The Curators Of The University Of Missouri Engineered biological nerve graft, fabrication and application thereof
US8852932B2 (en) 2004-02-24 2014-10-07 The Curators Of The University Of Missouri Self-assembling cell aggregates and methods of making engineered tissue using the same
US8931880B2 (en) 2010-10-21 2015-01-13 Organovo, Inc. Devices, systems, and methods for the fabrication of tissue
US9315043B2 (en) 2013-07-31 2016-04-19 Organovo, Inc. Automated devices, systems, and methods for the fabrication of tissue
US9332779B2 (en) 2014-02-05 2016-05-10 Modern Meadow, Inc. Dried food products formed from cultured muscle cells
US9481868B2 (en) 2014-10-06 2016-11-01 Organovo, Inc. Engineered renal tissues, arrays thereof, and methods of making the same
US9752122B2 (en) 2013-09-13 2017-09-05 Modern Meadow, Inc. Edible and animal-product-free microcarriers for engineered meat
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